薄层层析-图像分析定量热凝胶β-1，3-葡寡糖

单向两次薄层层析展开可以显著提高对热凝胶β-1，3-葡寡糖的分析效果，以Silica gel 60 F254硅胶板为固定相，V（乙酸）∶V（乙酸乙酯）∶V（水）=2∶2.4∶1.4为展开剂，展距20 cm时，单向两次展开可以较好地分离聚合度为2～13的热凝胶β-1，3-葡寡糖。此外还建立了Image J软件图像分析定量热凝胶β-1，3-葡寡糖的方法，在不同糖质量浓度范围（0.02～0.1、0.1～0.8、0.8～5.0 g/L）内寡糖显色斑点的灰度积分响应值与糖浓度分别有着很好的线性关系（R2＞0.99）。该方法的灵敏度高达0.02 g/L，相对标准偏差为1 %～5 %，稳定性很好。与HPLC等方法相比具有操作简便、分析快速、不需专门仪器和特定寡糖标准品等优点。 关键词： 热凝胶；β-1，3-葡寡糖；薄层层析；图像分析；Image J软件 中图分类号：TQ 281 文献标志码：A 文章编号：1673—1689（2016）11—1156—07
The study first found that a unidirectional-twice thin layer chromatography significantly improved the analysis of Curdlan β-1，3-glucooligosaccharides. With the best developing reagents acetic acid，ethyl acetate and H2O in the ratio of 2∶2.4∶1.4 on the Silica gel 60 F254 plate of 20 cm，Curdlan β-1，3-glucooligosaccharides could be separated from the degree of polymerization 2 to 13. Then，the Image J software was used to quantify Curdlan β-1，3-glucooligosaccharides. A good linear relationship（R2＞0.99） existed between saccharides concentration within three ranges of 0.02 g/L～0.1 g/L，0.1 g/L～0.8 g/L and 0.8 g/L～5.0 g/L and grayscale integration respond. The established quantification had a detection limit of 0.02 g/L and a relative standard deviation of 1 %～5 % with good stability. Compared with other methods such as HPLC，this method is simple and rapid with no specialized instruments and oligosaccharides standards